We examined the radial growth - climate association of a disjunct population of white spruce (Picea glauca (Moench) Voss) at its southern limit of distribution. Forty-four white spruce tree islands were sampled over four mixed-grass prairie preserves in the Spruce Woods Provincial Park located in the forestprairie boundary of southwestern Manitoba. Reduced radial growth occurred during the 1910s, 1930s, early 1960s, and the late 1970s to the early 1980s and corresponded to periods of drought on the Canadian prairies, and the Great Plains of the United States. Correlation and response function coefficients indicated that conditions in the summer and fall of the previous year (t-1), and the summer of the current year (t) strongly influenced white spruce growth. Growth was positively correlated with August-September (t-1) and May-June-July (t) precipitation and moisture index (precipitation minus potential evapotranspiration). Radial growth was positively associated with June-July-August (t) river discharge. Growth was most correlated with maximum and mean temperature compared with minimum temperature. Precipitation and maximum temperature accounted for the greatest variation in radial growth (61%). The results suggest that white spruce growth is sensitive to climatic fluctuations because growth is restricted by moisture deficiency exacerbated by temperature-induced drought stress.

The composition of lignin in the outer cell-wall layers of spruce and poplar has been studied and the data obtained have been compared with those of the mature reference wood in which the secondary cell wall predominates. Materials with exclusively or predominantly outer cell-wall layers were examined. Accurate data relating to the lignin monomer composition and the number of β-O-4´ bonds were obtained from pure middle lamella/primary cell wall lignin. Firstly, a 10 000 year old white spruce material, with most of the secondary cell wall missing, was studied. The aged lignin was composed of guaiacyl units only, and was slightly more condensed but otherwise similar to the reference lignin. Secondly, the developing xylem of a Norway spruce clone was analyzed during a growth season. In spring and early summer, growth is very rapid and the intention was to sample tissues in which the secondary cell-wall layers had not yet lignified, but where the outer layers at least had started to lignify. Microscopy, Klason lignin and carbohydrate analyses showed that the lignin in the developing xylem of samples from mid-June was located exclusively in the middle lamella. The lignin was more condensed, was composed of guaiacyl units only and contained more end-groups than the reference Norway spruce wood. Thirdly, the cambial tissues of a Balsam poplar clone were surveyed during a growth season. Both the phloem side and the xylem side of the cambial region were examined. The Klason lignin content and carbohydrate monomer distribution showed that in June and August the tissues on the phloem side contained material with mainly middle lamella/primary walls. In June, the xylem side in the cambial region contained mainly middle lamella/primary walls, and in August the secondary cell wall carbohydrates were being deposited. Both tissues contained lignin that was more condensed and had more end-groups than the reference lignin. In mid-June, the developing xylem had a ratio of syringyl to guaiacyl units of 0.6, whereas the ratio for the reference wood was 1.3. In the final study, lignin from the primary cell walls from a hybrid aspen cell suspension culture was investigated. The lignin contained only guaiacyl units which were more condensed than those observed in the reference poplar wood. / <p>QC 20100920</p>

En el marco del proyecto de investigación general "Diversidad arbórea en la zona mediterránea en Chile: ¿la silvicultura cercana a la naturaleza y la rentabilidad son mutuamente excluyentes?" en la VIII. Region del Biobio (desde 2018 la XVI. Región de Ñuble) se realizó un ensayo de campo con tres especies de árboles nativos chilenos en la zona mediterránea de Chile en el período de vegetación de 2012/2013. El centro de Chile se caracteriza por una alta biodiversidad con muchas especies endémicas. Dado que gran parte de los ecosistemas forestales naturales ya han sido destruidos, el establecimiento óptimo de bosques con especies de árboles nativos es particularmente importante y aún no se ha investigado lo suficiente. En esta tesis doctoral, se examinó cómo estas especies pueden ser (re-)establecidas en la zona central y qué condiciones ambientales influyen en su crecimiento y sus características ecofisiológicas. Se seleccionaron tres especies que son típicas de la región y en parte endémicas: dos especies deciduos de haya del sur Nothofagus obliqua (Roble) y Nothofagus glauca (Hualo) y una especie de hoja dura, Quillaja saponaria (Quillay). Estas especies fueron plantadas en la VIII. Región de Chile bajo tres coberturas diferentes: dentro de un bosque de Nothofagus glauca de 25 años previamente raleado, en una plantación joven de Pinus radiata, y en un área abierta con sombra lateral de bosques adyacentes más viejos de Pinus radiata y Nothofagus glauca. Para todas las plantas examinadas, se determinaron factores ambientales tales como la luz (factor de sitio indirecto), el contenido volumétrico de agua del suelo, la profundidad del suelo, la biomasa de la vegetación que rodea las plantas y la distancia al árbol vecino más cercano. La supervivencia, la longitud del brote y el diámetro del cuello de la raíz, la conductancia estomática, el área foliar específica y el número de las hojas de las plantas se determinaron como variables de respuesta. Con N. obliqua y Q. saponaria se fue un experimento de cámara climática en Alemania, no se pudo establecer N. glauca en el invernadero. Se estudiaron doce plantas por especie x 6 tratamientos en dos cámaras climáticas con las mismas condiciones de temperatura y luz. El crecimiento, la asignación de biomasa, el número de hojas, el área foliar específica, la conductancia estomática y la actividad fotosintética de las plantas se investigaron bajo dos niveles diferentes de luz y tres niveles diferentes de agua. Para el experimento en el campo, la pregunta de investigación fue qué intensidad de la luz y, por lo tanto, qué grado de cobertura del dosel es la opción correcta para el establecimiento de las tres especies de árboles mediante la plantación y si se puede determinar la disponibilidad óptima de luz. Las mortalidades más bajas se encontraron en el espacio abierto sombreado lateralmente y en el bosque de N. glauca. Se pudo demostrar que el crecimiento de las tres especies de árboles fue mayor en el espacio abierto con sombra lateral. Para N. obliqua, la disponibilidad de luz fue particularmente importante bajo dosel. Q. saponaria respondió con una disminución del crecimiento entre el gradiente de luz de 73% a 87%, de modo que un posible óptimo de luz para esta especie, que no podría determinarse a partir de este estudio debido a su diseño, varía de 64% a 71% ISF . Además, se preguntó al ensayo de campo si la disponibilidad de radiación explica suficientemente el crecimiento, la conductancia estomática y el área foliar específica de estas especies o si otros factores ambientales son importantes. Para las tres especies, se encontraron factores de influencia adicionales en diversos grados, por ejemplo humedad del suelo, profundidad del suelo y los efectos de la competencia de los bosques adultos y la vegetación herbácea circundante. También se podría mostrar una influencia de la fertilización previa. La tercera pregunta para el ensayo de campo fue qué fase dentro de la estación de crecimiento es crucial para el crecimiento de cada una de estas especies. Aquí se pudo demostrar que las tres especies hicieron la mayor contribución al crecimiento del brote entre octubre y enero. En la fase subsiguiente, el crecimiento se redujo drásticamente. Para el experimento de la cámara climática, se formularon preguntas de investigación sobre cómo los parámetros de crecimiento, la distribución de biomasa, el área foliar específica, el recuento foliar, la conductancia estomática y la fotosíntesis de Nothofagus obliqua y Quillaja saponaria responden a diferentes niveles de luz y estrés hídrico. Bajo estrés hídrico, el crecimiento, la biomasa total, la conductancia estomática y la actividad fotosintética se redujeron en ambas especies. En ambas especies, se observó una asignación de biomasa a las raíces al aumentar el estrés hídrico, pero mucho más pronunciada en N. obliqua que en Q. saponaria. Si bien Q. saponaria no cambió su asignación de biomasa de otra manera, N. obliqua redujo su fracción de hoja relativa. El área foliar específica de ambas especies fue menor en luz plena que en sombra. El recuento de hojas mostró tanto una reacción a la luz como el estrés hídrico. Fue menor bajo la sombra y con el aumento del estrés hídrico. Sin embargo, Q. saponaria mostró una adaptación más clara a los períodos de sequía manteniendo su número de hojas bajo sombra en todas las variantes de estrés hídrico. En una síntesis, los resultados de ambos experimentos fueron combinados y comparados. Las recomendaciones silvícolas se derivaron de los resultados combinados. Al establecer las tres especies de árboles en un corte tala rasa mas pequeño, debe garantizarse el sombreado lateral mediante bosques adyacentes. Opcionalmente, las especies de hoja dura como Q. saponaria pueden introducirse como plantas nodrizas. Para una conversión de plantaciones exóticas a bosques mas naturales manejados de manera sostenible, se recomiendan cortes de tala rasa de tamaño pequeño. Si las especies nativas se establecen bajo un bosque natural, el raleo regular es importante. Esto proporciona más luz y una mayor disponibilidad de recursos para los árboles jóvenes plantados. El establecimiento conjunto de Pinus radiata y las especies de árboles nativos solo se puede recomendar en una medida limitada debido al crecimiento más rápido de P. radiata y la competencia por los recursos que resulta.

The Arabidopsis chitinase-like1 (Atctl1) mutant, pom1 is compromised in primary cell wall development, resulting in short roots when grown on high sucrose and shortened hypocotyls when grown in darkness. To better understand this phenotype and the evolution of AtCTL1 and its homologue, AtCTL2, we obtained a large number of CTL sequences and determined the phylogenetic relationships among them. Since microarray analysis had suggested a change in auxin response or homeostasis in pom1, I used the auxin reporter DR5::GUS in the pom1 background to assess changes in distribution. To assess whether the biochemical functions of AtCTL1 homologues in Arabidopsis and other plants are conserved, I transformed pom1 with AtCTL2 and CTLs from poplar (Populus trichocarpa x Populus deltoides clone H-11) and from Picea glauca (spruce) and assessed rescue of the pom1 phenotype. To further understand CTL expression and function, Arabidopsis and poplar CTL promoter::GUS fusions were also expressed in Arabidopsis, PopCTL1 overexpressed in Arabidopsis, and CTL expression down regulated in poplar by RNAi. Our results indicate that CTL genes represent an ancient family encoding proteins of conserved biochemical function. In dicots, represented by Arabidopsis and poplar) duplicated CTL genes are differentially expressed in conjunction with primary and secondary cell wall development, respectively. Mutation of these genes results in improperly formed primary walls in certain cell types in the case of AtCTL1, and an impairment in the differentiation of vascular bundles for AtCTL2. Overexpression of PopCTL1 in Arabidopsis seems to over stimulate the differentiation of vascular bundles, and our studies show that auxin distribution is altered in the Atctl1 mutant. Down regulation of PopCTL1 and PopCTL2 in poplar appears to phenocopy aspects of these mutations, resulting in secondary cell walls that appear to have less deposition of lignin and an accelerated production of secondary xylem respectively. While specific biochemical function(s) of CTL genes were not studied, potential functions are discussed.

The Arabidopsis chitinase-like1 (Atctl1) mutant, pom1 is compromised in primary cell wall development, resulting in short roots when grown on high sucrose and shortened hypocotyls when grown in darkness. To better understand this phenotype and the evolution of AtCTL1 and its homologue, AtCTL2, we obtained a large number of CTL sequences and determined the phylogenetic relationships among them. Since microarray analysis had suggested a change in auxin response or homeostasis in pom1, I used the auxin reporter DR5::GUS in the pom1 background to assess changes in distribution. To assess whether the biochemical functions of AtCTL1 homologues in Arabidopsis and other plants are conserved, I transformed pom1 with AtCTL2 and CTLs from poplar (Populus trichocarpa x Populus deltoides clone H-11) and from Picea glauca (spruce) and assessed rescue of the pom1 phenotype. To further understand CTL expression and function, Arabidopsis and poplar CTL promoter::GUS fusions were also expressed in Arabidopsis, PopCTL1 overexpressed in Arabidopsis, and CTL expression down regulated in poplar by RNAi. Our results indicate that CTL genes represent an ancient family encoding proteins of conserved biochemical function. In dicots, represented by Arabidopsis and poplar) duplicated CTL genes are differentially expressed in conjunction with primary and secondary cell wall development, respectively. Mutation of these genes results in improperly formed primary walls in certain cell types in the case of AtCTL1, and an impairment in the differentiation of vascular bundles for AtCTL2. Overexpression of PopCTL1 in Arabidopsis seems to over stimulate the differentiation of vascular bundles, and our studies show that auxin distribution is altered in the Atctl1 mutant. Down regulation of PopCTL1 and PopCTL2 in poplar appears to phenocopy aspects of these mutations, resulting in secondary cell walls that appear to have less deposition of lignin and an accelerated production of secondary xylem respectively. While specific biochemical function(s) of CTL genes were not studied, potential functions are discussed.

The main task of this thesis was to investigate the influence of the preservation by ethyl alcohol and dry mounting on the morphometric characteristics of water bugs´ and to test the hypothesis that live individuals and individuals preserved by ethyl alcohol and dry mounted (both groups deal with individuals of the same species, sex and developmental stage) have the same morphometric parameters. The next task was to determine the effect of preparation by ethyl alcohol and dry mounting on the change of weight caused by different way of individuals´ preparation. Two model species of water bugs were chosen for this study: creeping water bug Ilyocoris cimicoides (Naucoridae) and backswimmer Notonecta glauca (Notonectidae). Weight and twelve other morphometric body characters were measured in adults of both sexes of both species. All data and their differences were tested by Student's paired t-test or nonparametric Wilcoxon test. The level of significance p = 0,05 was used for the decision whether the difference before and after preservation by ethyl alcohol and dry mounting is or is not statistically significant. Both preservation by ethyl alcohol and dry mounting influence only the body weight of treated individuals. Other morphometric characters are more or less independent on the way of preservation of water bugs´ individuals in collections. In this work, we also tested the effect of the Dyar rule for the increase in weight and size between different developmental stages of both species studied. Dyar rule for the evolution of both species was not verified. This project was supported by the grant of the Grant Agency of the Czech Republic No. P505/10/0096.

The aim of this study was to verify the possibility of ibuprofen degradation by selected plant cultures and determination of activities of antioxidant enzymes (peroxidase, catalase, ascorbate peroxidase and glutathione-S-transferase) as markers of oxidative stress caused by ibuprofen. Nicotiana tabaccum (cv. La Burley 21, cv. SR 1 and their GMOs) and Nicotiana glauca were used as experimental plants. The rate of removal of ibuprofen tested by tobacco was decreasing in the following order: N. tabaccum SR1 > N. tabaccum Zm-P60-1-T4 > N. tabaccum TRI 2T2 > N. glauca > N. tabaccum TRI 2T1 > N. tabaccum cv. La Burley > N. tabaccum Zm-P60-1-T5. As the most suitable tobacco for the removal of ibuprofen seemed untransformed N. tabaccum SR1. The long-term experiment showed that plant stress is being manifested even after longtime. N. tabaccum cv. La Burley 21 seemed to be the most tolerant to ibuprofen in compare with the total enzyme activities in cultures with the presence of ibuprofen and controls. N.glauca was the least tolerant cultivar. Keywords: phytoremediation, ibuprofen, Nicotiana tabaccum, Nicotiana glauca, HPLC, peroxidase, catalase, ascorbate peroxidase, glutathion-S-transferase

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